LOCUS       sequence-478811-        7509 bp    DNA     circular SYN 16-JUL-2025
DEFINITION  Expresses EGFP-NUP98 fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7509)
  AUTHORS   Zhang H, Lesnov GD, Subach OM, Zhang W, Kuzmicheva TP, Vlaskina AV, 
            Samygina VR, Chen L, Ye X, Nikolaeva AY, Gabdulkhakov A, Papadaki S,
            Qin W, Borshchevskiy V, Perfilov MM, Gavrikov AS, Drobizhev M, 
            Mishin AS, Piatkevich KD, Subach FV
  TITLE     Bright and stable monomeric green fluorescent protein derived from 
            StayGold.
  JOURNAL   Nat Methods. 2024 Apr;21(4):657-665. doi: 
            10.1038/s41592-024-02203-y. Epub 2024 Feb 26.
  PUBMED    38409224
REFERENCE   2  (bases 1 to 7509)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41592-024-02203-y"; journalName: "Nat Methods"; date: 
            "2024-04"; volume: "21"; issue: "4"; pages: "657-665"
COMMENT     Sequence Label: pEGFP-NUP98
FEATURES             Location/Qualifiers
     source          1..7509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4297..4418
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4334..4353)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4388..4407
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4425..4880)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4562..4583)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4774..4793
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4907..5011
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5013..5370
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5013..5204
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5036..5056)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5221..5356
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5405..6199
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5405..6199
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5459..5478)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6069..6088
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6387..6406)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6431..6478
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6807..7395
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7296..7315
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggccggac tcagatctcg agcatgttta acaaatcatt tggaacaccc
     1381 tttgggggtg gcacaggtgg ctttggcaca acttcaacat ttggacagaa tactggcttt
     1441 ggcactacta gtggaggggc atttggaaca tctgcatttg gttctagcaa caatactgga
     1501 ggcctctttg gaaattcaca gactaaacca ggaggattgt ttggaaccag ttcatttagc
     1561 cagccagcta cctccacaag cactggcttt gggtttggta cgtcaacagg aacagcaaat
     1621 accttgtttg gaactgcaag cacagggacc agtctcttct catcccaaaa caatgccttt
     1681 gcacaaaata aaccaactgg ctttggcaat tttggaacca gtactagcag tggaggactc
     1741 tttggaacca caaataccac ctctaatcct tttggcagca catctggctc cctctttggg
     1801 ccaagtagtt ttacagctgc tcctactggg actactatta aatttaaccc tccaactggt
     1861 acagatacta tggtcaaagc tggagttagc actaacataa gtaccaagca ccagtgtatt
     1921 actgctatga aagaatatga aagcaagtca ctagaggaac ttcgtttaga ggattatcag
     1981 gctaacagga agggcccaca gaaccaggtg ggagcaggta ccacaactgg cttgtttggg
     2041 tcttctccag ccacttccag cgcaacagga ctcttcagct cctccaccac taattcaggc
     2101 tttgcatatg gtcagaacaa aactgccttt ggaactagta caactggatt tggaacaaat
     2161 ccaggtggtc tctttggcca acagaatcag cagactacca gcctcttcag caaaccattt
     2221 ggccaggcta caaccaccca gaacactggc ttttcctttg gtaataccag caccatagga
     2281 cagccaagca ccaacaccat gggattattt ggagtaaccc aagcctcaca gcctggaggt
     2341 ctttttggga cagctacaaa caccagcact gggacagcat ttggaacagg aacaggtctc
     2401 tttgggcaga ccaatactgg atttggtgct gttggttcga ccctgtttgg caataacaag
     2461 cttactacat ttggaagcag cacaaccagt gcaccttcat ttggtacaac cagtggcggg
     2521 ctctttggta acaaaccaac cctgacttta ggaaccaata caaacacttc taattttggt
     2581 tttggcacaa ataccagtgg gaatagtatt tttggaagta aaccagcacc tgggactctt
     2641 ggaactgggc ttggtgcagg atttggaaca gctcttggtg ctggacaggc atctttgttt
     2701 gggaacaacc aacctaagat tggagggcct cttggtacag gagcctttgg ggcccctgga
     2761 tttaatacta cgacagccac tttgggcttt ggagcccccc aggccccagt agctttgaca
     2821 gatccaaatg cttctgctgc ccagcaggct gttctccagc agcacatcaa tagtctaaca
     2881 tactcacctt ttggagactc tcctctcttc cggaatccga tgtcagaccc taagaagaag
     2941 gaagagagat tgaaaccaac aaatccagca gcccagaagg ctcttactac acctactcat
     3001 tataaactga caccccgccc tgccactaga gtccggccaa aggctttaca aacaacaggc
     3061 acagccaagt cacatctctt tgatgggctg gatgacgatg aaccatccct agccaatgga
     3121 gcattcatgc ccaagaagag cattaagaag ttggttttga agaaccttaa taatagcaat
     3181 ctcttttctc ctgttaatcg tgattcagaa aatctagctt caccatctga atatccagaa
     3241 aatggagaga gatttagttt cctaagcaaa cctgttgatg agaatcacca gcaggatgga
     3301 gatgaagatt cccttgtttc acatttttat actaacccta ttgccaaacc tattcctcaa
     3361 accccagaaa gtgctggaaa taaacacagc aacagcaaca gtgtggatga taccattgtt
     3421 gcattaaaca tgcgtgctgc tttgcgaaat gggctggaag gaagcagtga agaaacgtct
     3481 tttcatgatg agtcacttca ggatgaccga gaagaaatag aaaataattc ttaccatatg
     3541 cacccagcag gtattattct cactaaggtt ggttactata ctattccatc tatggatgac
     3601 cttgctaaaa ttaccaatga aaaaggagag tgcattgtct ctgatttcac tattggtcgg
     3661 aaaggttatg gttcaatcta ttttgaagga gatgtgaatt tgacaaatct aaatttggat
     3721 gatattgtgc atatccggag gaaagaagta gttgtctact tagatgataa ccaaaaacca
     3781 cctgtgggtg aagggctaaa taggaaggct gaagttacat tggatggagt ttggccaaca
     3841 gataaaacat ctcgttgttt aataaagagc ccagatcgcc ttgctgatat caactatgaa
     3901 ggaagattgg aagcagtttc aaggaaacag ggagctcaat tcaaagaata ccggcctgaa
     3961 actggttctt gggtgtttaa ggtctcccat ttttctaagt atggccttca ggattctgat
     4021 gaagaggagg aggagcatcc gtctaaaact agtacaaaga agttgaagac tgctcctttg
     4081 cctcctgcaa gccagactac gcccttgcag atggctctta atggcaaacc tgcacctcca
     4141 cctcaggtag agaaaaaagg acagtgagga tccaccggat ctagataact gatcataatc
     4201 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     4261 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     4321 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4381 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     4441 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4501 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4561 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4621 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4681 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4741 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     4801 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     4861 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4921 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4981 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     5041 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5101 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5161 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5221 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5281 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     5341 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     5401 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5461 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5521 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5581 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5641 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5701 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5761 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5821 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5881 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     5941 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     6001 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     6061 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     6121 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     6181 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     6241 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     6301 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6361 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     6421 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     6481 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     6541 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     6601 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     6661 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6721 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6781 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6841 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6901 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6961 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7021 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7081 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7141 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7201 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7261 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7321 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7381 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7441 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7501 gccatgcat
//
